Winters T A, Williams M V
Department of Medical Microbiology and Immunology, Ohio State University, Columbus 43210.
Virology. 1993 Aug;195(2):315-26. doi: 10.1006/viro.1993.1382.
The herpes simplex virus type 2 (HSV-2) uracil-DNA glycosylase (UNG) was from the nuclei of infected KB cells using ion exchange, affinity, and chromatofocusing chromatography techniques. Chromatography on DNA cellulose revealed two distinct HSV-2-encoded UNGs. One species, designated A, was purified approximately 324-fold, while the second species, designated B, was purified approximately 130-fold. The HSV UNG species B was observed to unidirectionally convert to the A species, suggesting that the B species binding to DNA cellulose may be the result of an association with other DNA binding proteins. SDS-PAGE demonstrated that both species A and B had molecular weights of 32,000. The HSV-2-encoded UNGs could be distinguished from the cellular nuclear UNG based upon differences in their behavior on the chromatography matrixes and by their molecular weights. There were no significant differences in the biochemical properties of the HSV-2-encoded or nuclear UNGs. Furthermore, all of these UNGs reacted with a monoclonal antibody produced against the human placental UNG. These data support recent studies, at both the DNA and the amino acid levels, which have demonstrated that this enzyme is highly conserved between species.
利用离子交换、亲和及聚焦层析技术，从感染的KB细胞核中分离出单纯疱疹病毒2型（HSV - 2）尿嘧啶DNA糖基化酶（UNG）。DNA纤维素柱层析显示出两种不同的由HSV - 2编码的UNG。一种命名为A，纯化倍数约为324倍；另一种命名为B，纯化倍数约为130倍。观察到HSV UNG的B型单向转化为A型，这表明与DNA纤维素结合的B型可能是与其他DNA结合蛋白缔合的结果。SDS - PAGE显示A、B两种类型的分子量均为32,000。基于它们在层析基质上行为的差异及其分子量，可将HSV - 2编码的UNG与细胞核UNG区分开来。HSV - 2编码的UNG与细胞核UNG的生化特性没有显著差异。此外，所有这些UNG均与针对人胎盘UNG产生的单克隆抗体发生反应。这些数据支持了最近在DNA和氨基酸水平上的研究，这些研究表明该酶在物种间高度保守。
